Respiratory Tree Research Articles

4-Hydroxyphenylpyruvate dioxygenase from sea cucumber Apostichopus japonicus negatively regulates reactive oxygen species production

As a wide distribution molecule, 4-hydroxyphenylpyruvate dioxygenase (4-HPPD) catalyzes the second step in the tyrosine catabolism pathway. This process commonly occurs in all aerobic life forms. The broad distribution of these metabolites suggests that they have an important role in many organisms. A portion of the 4-HPPD homology sequence was also identified in Apostichopus japonicus transcriptome. However, the functional roles of A. japonicus 4-HPPD remain unclear. In the current study, a 4-HPPD homolog was cloned from A. japonicus (designated as AjHPPD). The nucleotide sequence analysis showed that the open reading frame of AjHPPD was 1149 bp and encoded a 382-amino-acid residue polyprotein with glyoxalase_4 (residues 20–133) and glyoxalase (residues 180–335) domains. The spatial expression analysis revealed that AjHPPD was ubiquitously expressed in all examined tissues with large-magnitude in the respiratory tree and was minimally expressed in coelomocytes. Compared with a control group, the significant increase in transcription of AjHPPD mRNA in the Vibrio splendidus-challenged sea cucumber was 2.10-fold (p < 0.01) at 48 h and returned to the normal level at 72 and 96 h. Similarly, compared with a control group, the significant increase in the transcription of AjHPPD mRNA was 3.36-fold (p < 0.01) at 24 h after stimulation with 10 mg mL−1 of LPS. On the one hand, silencing AjHPPD in vitro could inhibit the expression of pentose phosphate pathway (PPP) flux enzyme glucose-6-phosphate dehydrogenase (G6PD) at the mRNA level and prevent the clearance of reactive oxygen species (ROS) in sea cucumbers. On the other hand, interference of AjHPPD by using specific siRNA can result in the significant promotion of coelomocyte apoptosis with a 1.61-fold increase in vitro. AjHPPD negatively regulated ROS levels by modulating tyrosine catabolism on AjG6PD expression and coelomocyte apoptosis in response to pathogen infection.

Effects of aerial exposure on oxidative stress, antioxidant and non-specific immune responses of juvenile sea cucumber Apostichopus japonicus under low temperature

Desiccation is a commonly stressful situation experienced by sea cucumber during transportation without/less water. The present study was conducted to investigate the effects of aerial exposure on the survival, oxidative damage, antioxidant capacity, immune-related response and gene expression of Apostichopus japonicus at different low temperatures. After acclimation, sea cucumbers were randomly divided into 3 groups, which were exposed to 5 °C, 10 °C and 15 °C in the closed laboratory condition, respectively. Each group has three parallel replicates. During the experiment, coelomic fluid and respiratory tree of A. japonicus were sampled at the time points of 0, 3, 6, 12, 24 and 48 h post-desiccation for further analysis. The results showed that the survival rates of sea cucumber significantly decreased as time prolonged, and those of 5 °C at 6–48 h of desiccation were significantly higher than 15 °C. Most oxidative damage parameters (e.g., O2− production, MDA, LPO and PC contents) significant increased after 6–12 h of desiccation. Antioxidant enzyme activities and T-AOC in coelomic fluid firstly increased and then decreased during aerial exposure, indicating that sea cucumber could adjust antioxidant defense to reduce the concentrations of ROS and MDA as a strategy for protecting organisms from oxidative damage in the early stage (0–6 h) of desiccation. The relative expression levels of Hsp90 and Hsp70 mRNA in respiratory tree of sea cucumber exhibited similar rise-fall trends with antioxidant parameters, while immune enzyme activities of ACP, AKP, LSZ and T-NOS, and gene expression of TLR, Rel and p105 all significantly decreased as time prolonged. Overall, low temperature postponed the process of ROS formation and the depression of antioxidant and non-specific immune responses of sea cucumber within a certain extent, which implied that it might play a positive role in improvement of desiccation tolerance. This study not only contribute to better understand the adaption mechanisms of A. japonicus to desiccation stress, but also provide valuable information for sea cucumber culture and transportation.

Green fluorescent protein-tagged Vibrio splendidus for monitoring bacterial infection in the sea cucumber Apostichopus japonicus

Vibrio splendidus is an important opportunistic pathogen that infects many important marine animals; for example, it can cause skin ulcer syndrome in the sea cucumber Apostichopus japonicus. In this study, a green fluorescent protein (GFP)-tagged V. splendidus was constructed by ligating GFP into the plasmid pET28a. The growth of GFP-tagged V. splendidus showed no difference from that of the wild type strain. The distribution of V. splendidus in various tissues after artificial infection was determined by colony counting. When V. splendidus infected A. japonicus at a concentration of 5.0 × 107 CFU·mL−1, V. splendidus could be detected in the tissues of respiratory tree, intestine, body wall, tentacle, and muscle, with a level over 1.0 × 106 CFU·g−1 at 48 h post-infection and with V. splendidus remaining in coelomic fluid at a level of 2.1 × 109 CFU·mL−1. Green fluorescence was only observed in the respiratory tree and intestine in a paraffin section at 48 h of infection, which indicated that V. splendidus could adhere to these two tissues. Moreover, V. splendidus could adhere to coelomocytes of A. japonicus but it could poorly internalize coelomocytes in vitro. The adherence of V. splendidus to coelomocyte was dependent on the β-integrin on the surface of coelomocytes.

A tandem-repeat galectin-1 from Apostichopus japonicus with broad PAMP recognition pattern and antibacterial activity

Galectins belong to the family of carbohydrate-binding proteins and play major roles in the immune and inflammatory responses of both vertebrates and invertebrates. In the present study, one novel galectin-1 protein named AjGal-1 was identified from Apostichopus japonicas with an open reading frame of 1179 bp encoding a polypeptide of 392 amino acids. The deduced amino acids sequence of AjGal-1 contained three carbohydrate recognition domains (CRDs) which shared 34–37% identity with that of other galectin proteins from echinodermata, fishes, and birds. In the phylogenetic tree, AjGal-1 was closely clustered with galectins from Mesocentrotus nudus and Paracentrotus lividus. The mRNA transcripts of AjGal-1 were ubiquitously expressed in all the detected tissues, including gut, longitudinal muscle, gonad, coelomocytes, respiratory tree, tentacle and body wall, with the highest expression level in coelomocytes. After Vibrio splendidus stimulation, the mRNA expression levels of AjGal-1 in coelomocytes were significantly increased at 6 and 12 h (P < 0.01) compared with that in control group, and went back to normal level at 72 h. The recombinant protein of AjGal-1 (rAjGal-1) could bind various PAMPs including d-galactose, lipopolysaccharide (LPS), peptidoglycan (PGN) and mannose (Man), and exhibited the highest affinity to d-galactose. Meanwhile, rAjGal-1 could also bind and agglutinate different kinds of microorganisms, including gram-negative bacteria (V. splendidus and Escherichia coli), gram-positive bacteria (Micrococus leteus), and fungi (Pichia pastoris). rAjGal-1 also exhibited anti-microbial activity against V. splendidus and E. coli. All these results suggested that AjGal-1 could function as an important PRR with broad spectrum of microbial recognition and anti-microbial activity against the invading pathogen in A. japonicas.

Antifouling and antibacterial activities of bioactive extracts from different organs of the sea cucumber Holothuria leucospilota

In this study, antifouling and antibacterial properties of polar, semi polar and non-polar bioactive compounds derived from the sea cucumber Holothuria leucospilota were investigated. A series comprising n-hexane, ethyl acetate and methanol extracts from four different organs of sea cucumber (body wall, gonad, digestive tract and respiratory tree) were investigated for their antibacterial and antifouling activity against two species of microalgae, larvae of barnacle Amphibalanus amphitrite and a non-target organism: larvae of Artemia salina. Results showed that the highest antibacterial activity was found in the ethyl acetate extract of body wall against Staphylococcus aureus with minimum inhibitory concentration (MIC) of 0.250 mg/mL. The MIC of ethyl acetate extract of body wall ranged between 0.062 and 0.250 mg/mL against two microalgal strains. All extracts showed moderate to low toxicity against larvae of barnacles. The most pronounced anti-barnacle activity was found in the ethyl acetate extract of body wall with median lethal concentration (LC50) value of 0.049 mg/mL. Among the twelve extracts, the ethyl acetate extracts of body wall showed the maximum antibacterial and antifouling activities. The bioactive compounds of the ethyl acetate extract of the sea cucumber body wall were determined using gas chromatography coupled with mass spectrometry. Fatty acids and terpenes were the major compounds, which may be responsible for antibacterial and antifouling activity of the ethyl acetate extract of body wall of the sea cucumber H. leucospilota.

Pneumonia prevention in the elderly patients: the other sides.

Pneumonia is one of the leading causes of morbidity and mortality from infection in elderly patients. The increased frequency of pneumonia among elderly subjects can be explained by the physiological changes linked to the progressive aging of the respiratory tree and the diminished immunological response. A spiral of event leads to frailty, infection and possible death; preventing pneumonia consists of controlling the risk factors. Dysphagia, which is associated with malnutrition and dehydration, is recognized as one of the major pathophysiological mechanism leading to pneumonia and its screening is crucial for the pneumonia risk assessment. The impairment in the oropharyngeal reflexes results in stagnation of foreign material in the lateral cavities of the pharynx which may then get aspirated repeatedly in the lungs and cause pneumonia. Pneumonia prevention starts with lifestyle modifications such as alcohol and tobacco cessation. A careful review of the risk-benefit of the prescribed medication is critical and adaptation may be required in elders with multiple morbidities. Respiratory physiotherapy and mobilization improve the functional status and hence may help reduce the risk of pneumonia. Maintaining teeth and masticatory efficiency is important if malnutrition and its consequences are to be avoided. Daily oral hygiene and regular professional removal of oral biofilm can prevent the onset of periodontitis and can avoid an oral environment favoring the colonization of respiratory pathogens than can then be aspirated into the lungs.

Microplastic fibers transfer from the water to the internal fluid of the sea cucumber Apostichopus japonicus

Microplastics (MPs) are small plastic particles less than 5 mm in diameter. MPs in the form of microfibers (MFs) are widely detected in aquatic habitats and are of high environmental concern. Despite many reports on the effects of MFs on marine animals, their effect on sea cucumbers is still unclear. In addition, our previous filed study has shown that MFs may transfer to the coelomic fluid of the sea cucumber Apostichopus japonicus (A. japonicus). Here, we show how MFs transfer to the coelomic fluid of the sea cucumber. We captured the MFs during their transfer from the water to the coelomic fluid through the respiratory tree. A. japonicus ingested in the MFs along with the water during respiration; the MFs got stuck in the respiratory tree or transferred to the coelomic fluid. The transferred MFs increased during 72 h of exposure and persisted for 72 h after the transfer to clean water. Among the immunity indices, lysozyme (LZM) levels increased in response to the transferred MFs, which confirms the defensive role of LZMs against strange substances. Additionally, non-significantly decreased levels of total antioxidant capacity (T-AOC), malondialdehyde (MDA), peroxidase (POD) and phenol oxidase (PPO) were observed at 24 h and 48 h post-exposure, suggesting minimal oxidative imbalance. Furthermore, there were no significant changes in the speed and the total distance moved by A. japonicus post MFs transfer. This study revealed that MFs transfer and accumulate in the coelomic fluid of A. japonicus.

1520. Streptococcus anginosus Group Organisms Are an Increasing Cause of Complicated Sinusitis and Otitis Media in Children

BackgroundThe Streptococcus anginosus group (SAG), including S. anginosus, S. intermedius and S. constellatus, are common flora of the oral cavity, respiratory tree and gastrointestinal tract. However, these organisms have the potential to cause serious invasive infections and are notably pyogenic. We observed an apparent increase in the frequency of intraorbital and intracranial infections resulting from SAG at Texas Children’s Hospital (TCH). We undertook a retrospective review to describe the frequency and clinical features of these infections.MethodsWe reviewed the database of the clinical microbiology laboratory at TCH from 2011 to 2018 for SAG-positive cultures. For purposes of this study, cases included were those associated with 1) either otitis media or sinusitis and 2) Pott’s Puffy Tumor, orbital abscesses, epidural abscesses, subdural empyema, brain parenchymal abscesses, dural enhancement by imaging or mastoiditis. Similar methods were used for SAG identification throughout the study period. The number of cases per year was used along with annual hospital admissions data to estimate case rate; case rate trends were examined using linear regression.Results950 cultures positive for SAG were identified by the clinical lab; 95 cases met inclusion criteria. The median age of patients was 11.4 years. Specific diagnoses are presented in Figure 1. S. intermedius was most commonly isolated (81.1%) followed by S. constellatus (12.6%) and S. anginosus (7.4%); 50.5% of cases were polymicrobial. Among polymicrobial cases, S. aureus was most frequently isolated (25%). All patients underwent surgical intervention and 20.5% underwent ≥2 procedures (Figure 2). 16.8% were associated with intracranial thromboses and 4.2% with CNS infarcts; 8.4% of patients experienced persistent neurologic deficits. All isolates were susceptible to penicillin. We observed an increase in the annual disease rate during the study (Figure 3, P = 0.01).ConclusionComplications of otitis media and sinusitis due to SAG are associated with substantial morbidity. These infections are becomingly increasingly common at our center although the precise reason for this temporal trend is unclear. Multicenter studies are needed to validate these epidemiologic findings. Disclosures All authors: No reported disclosures.

1775 Tracheoesophageal Fistula Managed With Double Stenting of Bronchus and Esophagus

INTRODUCTION: Malignancy is responsible for 5-15% of acquired tracheoesophageal fistulas (TEF). Such patients have a short survival and management of these fistulas is a challenge. We present a case of TEF secondary to esophageal malignancy that was managed with double stenting of the bronchus and esophagus. CASE DESCRIPTION/METHODS: The patient is a 56 year old black female with no known past medical history who presented to our emergency department with complaints of chest pain, shortness of breath and a productive blood tinged cough. She had been recently discharged from another facility for a diagnosis of bronchitis, however symptoms had not abated. She reported a fifty-pound weight loss with poor oral intake. Her vital signs and labs on arrival were stable. Initial imaging in the ER was consistent with pneumonia of the right middle and lower lobes. She underwent a CT Neck, which showed esophageal wall thickening and a mass suspicious for an esophageal neoplasm. Due to concern of her recurrence of pneumonia and esophageal mass, she underwent an esophagram, which showed a stricture of the middle third of the esophagus with a fistula from the esophageal tumor to the right lower lobe bronchus. Due to concern of airway collapse with upper endoscopy, Pulmonology was consulted. The patient first underwent a bronchoscopy in which a Novatech Y-stent was deployed stabilizing the airway. The patient then underwent an endoscopy in which an esophageal stent was placed. Both procedures were successful and she tolerated oral intake well thereafter. She was discharged in stable condition to palliative hospice services. DISCUSSION: The case has relevance from both an interesting initial clinical presentation as well as management of this patient from both a technical and procedural aspect. This patient had continual infections from continuous overspill of esophageal contents into the respiratory tree. Although her prognosis to begin with was quite poor, these interventions allowed the patient some quality of life. This case also commands procedural respect. The patient’s left mainstem bronchus was extremely friable and actually ruptured during the initial course of balloon dilation. The stent in position did not completely cover the TEF however it did keep the central airway open. A long esophageal stent was placed by gastroenterology, which did completely cover the fistula. This patient’s case highlights potential difficulties in double stenting and anticipatory management by utilization of other services.

Cancer Progress and Priorities: Lung Cancer.

In the United States, lung cancer is the second most common diagnosed cancer and the leading cause of cancer-related death. Although tobacco smoking is the major risk factor accounting for 80% to 90% of all lung cancer diagnoses, there are numerous other risk factors that have been identified as

Can Different Body Tissues of Two Sea Cucumbers Supply a Fair Amount of Omega 3 for Health Benefit?

ABSTRACTThe lipid content and fatty acids profile of different tissues of Holothuria tubulosa and H. polii were analyzed to assess the suitability of the central Mediterranean species as a potential marine resource for fatty acids, in particular omega-3 polyunsaturated fatty acids (n-3 PUFAs). Simultaneously, their important indices for human health were evaluated. Gonad and respiratory tree had the highest lipid content, while the body had the lowest values. The results showed significant differences between species and among the different portions analyzed. Internal tunic and longitudinal muscle of H. tubulosa exhibited the highest PUFA proportion (55.3 and 46.4%, respectively). Eicosapentaenoic acid (EPA, C20:5n-3) and arachidonic acid (ARA, C20:4n-6) were the most abundant PUFAs in both species, with a favorable n-3/n-6 ratio in all tissues. The atherogenic and thrombogenicity indices and hypocholesterolaemic/hypercholesterolaemic fatty acid ratio suggested the high-quality of this food, similar to the nutritional values of most popular fish.

A novel caspase-1 mediates inflammatory responses and pyroptosis in sea cucumber Apostichopus japonicus

Caspase-1 is the most important inflammatory caspase and plays necessary roles in cell death. In the current study, a novel caspase-1 homolog was identified from sea cucumber Apostichopus japonicus; this homolog encoded a predicted protein containing a CASc domain with the pentapetide active site 173QACRG177. Ajcaspase-1 mRNA was ubiquitously expressed in all detected tissues, with higher levels found in respiratory trees and coelomocytes. Subcellular localization of Ajcaspase-1 mainly occurred in the nucleus. The transcript expression of Ajcaspase-1 was remarkably up-regulated in coelomocytes after Vibrio splendidus challenge in vivo and LPS stimulation in vitro. Recombinant Ajcaspase-1 (rAjcaspase-1) and the C-terminal of Ajcaspase-1 (rAjcaspase-1-C) revealed high levels of LPS binding activity and exhibited proteolytic activities toward the substrates Ac-YVAD-pNA and Ac-DEVD-pNA. The high activity of these enzymes in Ac-YVAD-pNA suggests that Ajcaspase-1 shares common caspase-1 activity. No obvious activities were detected for rAjcaspase-1-N. More importantly, rAjcaspase-1 significantly increased the expressions of HomoIL-1β and HomoGSDMD mRNA in HEK293 cells. Our results clearly indicate that Ajcaspase-1 may participate in the pyroptosis pathway by regulating inflammatory responses.

Identification and sequencing of the gene encoding DNA methyltransferase 3 (DNMT3) from sea cucumber, Apostichopus japonicus.

The sea cucumber Apostichopus japonicus is well known as a traditional tonic food and as a commercially important cultured aquatic species. This species produces saponins, and has a high potential to cope with environmental stress, such as aestivation, organ regeneration, and wound healing. Recently, several studies have shown that cellular reprogramming and the physiological responses of the sea cucumber to environmental changes, including aestivation, are potentially mediated by epigenetic DNA methylation. The DNA methyltransferase (DNMT)1 and DNMT3 genes are independent participants in the maintenance and de novo methylation of specific sequences. Sea urchin (Strongylocentrotus purpuratus) and starfish (Asterina pectinifera), which belong to the same phylum as A. japonicus, have both DNMT1 and DNMT3 genes. However, it was previously reported that DNMT1 is present, but DNMT3 is absent, in A. japonicus. In the present study, we sequenced the full-length cDNA of the A. japonicus DNMT3 gene. The newly sequenced DNMT3 gene comprises three major conserved domains (Pro-Trp-Trp-Pro (PWWP), plant homeodomain (PHD), and S-adenosylmethionine-dependent methyltransferase (AdoMet-MTase)), indicating that the DNMT3 possibly has de novo DNA methylation catalytic activity. Gene structure and phylogenetic analysis showed that sea cucumber DNMT3 is evolutionarily conserved in the Echinodermata. Next, we demonstrated the conservation of DNMT3 gene expression in sea cucumber and starfish belong to same phylum, echinoderm. Using reverse transcription-polymerase chain reaction, sea cucumber DNMT3 mRNA was detected in testis tissue, but not in other tissues tested, including the respiratory tree, muscle, tentacle, intestine, and ovary. This is inconsistent with previous reports, which showed the expression of DNMT3 in ovary, but not in testis of the starfish A. pectinifera, indicating the tissue- and species-specific expression of DNMT3 gene. Although further studies are needed to clarify the epigenetic regulatory mechanisms of DNMT3 and its application to the aquaculture industry, our findings may provide insights into the sea cucumber biology.

The distribution, expression of the Cu/Zn superoxide dismutase in Apostichopus japonicus and its function for sea cucumber immunity

Cu/Zn superoxide dismutases (SODs) are antioxidative metalloenzymes that exist ubiquitously in different species and are distributed widely in various tissues and cell types. In this study, the distribution and biological function of the Cu/Zn superoxide dismutase in Apostichopus japonicus (AjSOD1) is first characterized. The AjSOD1 cDNA is 1219 bp in length and contains an open reading frame (ORF) of 459 bp that encodes a protein of 152 amino acids with a deduced molecular weight of 15.47 kDa and a predicted isoelectric point of 5.65. The Cu2+/Zn2+ binding domain and conserved residues were found in the AjSOD1 amino acid sequence. A quantitative reverse transcriptase real-time PCR (qRT-PCR) assay was developed to assess the expression of AjSOD1 in different tissues. Spatial distribution analysis showed that AjSOD1 was constitutively expressed in all tested tissues, with strong expression in the intestine and weak expression in the respiratory tree. mRNA Expression of AjSOD1 was significantly upregulated when challenged with the pathogen Vibrio splendidus. Functional investigation revealed that recombinant AjSOD1 displayed good antioxidant activity. More importantly, the addition of AjSOD1 resulted in a significant decrease in coelomocyte apoptosis by LPS/H2O2 challenge in vitro. The results indicate that sea cucumber SOD1 may play critical roles not only in the defense against oxidative stress but also in the innate immune defense against bacterial infections.

Congenital Lung Malformations.

Congenital lung malformations (CLM) comprise a spectrum of anatomical anomalies of the lungs and respiratory tree. The prenatal growth pattern of CLMs is unpredictable with larger lesions causing life-threatening complications, such as hydrops fetalis, and smaller lesions remaining asymptomatic and potentially regressing. The most common CLMs are congenital pulmonary adenomatoid malformations, bronchopulmonary sequestrations, congenital lobar emphysema, and bronchogenic cysts. All these lesions have clinically similar presentations when symptomatic, but pathophysiologic differences that must be considered when evaluating and treating. This article reviews the most common CLMs, their pathophysiology, clinical presentation, diagnostic considerations, and current literature on the controversies surrounding CLM management. [Pediatr Ann. 2019;48(4):e169-e174.].

Tracheobronchopathia Osteochondroplastica: A Case Report

Tracheobronchopathia osteochondroplastica (TOCP) is a rare disease defined by the presence of osteocartilaginous foci in the sub mucosa, without direct connection to the cartilage structures of the respiratory tree.

ROS-1 re-arrangements and c-MET amplifications in adenocarcinoma lung: A tertiary care center study from North India.

C-ros oncogene 1, receptor tyrosine kinase (ROS 1) proto-oncogene 1, receptor tyrosine kinase (ROS-1) fusions are potent oncogenic drivers and these re-arrangements promote signal transduction programs leading to uninhibited cell survival and proliferation identified in 1-2% of cases of nonsmall-cell lung cancer. Mesenchymal epithelial transition factor (MET) receptor tyrosine kinase and its ligand are predominantly involved in epithelial mesenchymal transition and tissue regeneration. The MET amplification and overexpression is oncogenic in 3-7% cases. The objectives of this study were to identify the frequency of ROS-1 and c-MET protein expression in adenocarcinoma lung and to correlate it with the clinicopathological parameters and to analyze the histomorphology of cases that harbor the characteristic mutations (c-MET and ROS-1). Study group comprised a prospective cases series of 90 cases of adenocarcinoma lung. ROS-1 protein expression was determined by immunohistochemistry using the D4D6 rabbit monoclonal antibody (Cell Signaling, Danvers, MA) and c-MET protein expressed was analyzed using the SP-44 clone (Ventana Medical Systems). c-MET protein expression was identified in 33.33% cases (n = 30/90) with statistically significant thyroid transcription factor-1 (TTF-1) positivity. ROS-1 protein expression was detected in 3.33% cases (n-3/90), in biopsies from the respiratory tree with TTF-1 expression. This is the first study from the Indian subcontinent to identify the frequency of ROS-1 re-arrangements and MET amplification in the Indian population. The availability of targeted therapy that has a significant impact on survival makes it essential to detect these less frequent mutations.

Journal Club-Electronic Cigarettes and Vaping as a Harm Reduction Alternative: Really?

Journal Club-Electronic Cigarettes and Vaping as a Harm Reduction Alternative: Really?

Effect of Vaccination on Influenza Infection Rate on KSA

Influenza is an acute respiratory disease characterized in its full form by the sudden onset of high fever, coryza, cough, headache, malaise, and inflammation of the upper respiratory tree and trachea. In most cases, pneumonic involvement is not clinically prominent. Aim of study: this Review, we examine the influenza virus infection in cross section study in Riyadh city KSA, This knowledge is used for risk assessments for future, and study effect of influenza vaccine and its implication against influenza virus infection rate STUDY METHODS: A cluster randomized trial involving 1271 children and adolescents and adult, age group between 36 months to 60 years who attend for SMC (Specialized medical center hospital) hospital in ER from January 1 2018, and ended December 31, 2018. History was taken for received vaccine or not, and risk factor diseases (Bronchial Asthma, Diabetics, Heart disease, pneumonia, Immunocompromised). NASOPHARNGEAL swab for: Diagnostic tests: rapid antigen testing, reverse transcription polymerase chain reaction (RT-PCR), and rapid molecular assays, and Follow-up for outcome in infected cases Results: the study revealed that 361of 1271 (28.4%) had influenza, 89 patient had H1N1 (7%), 128 patients had influenza A (10.07%) & 144 patients had influenza B (11.32%), one case had H1N1 and Influnza B Detected, 61% had received vaccination., Among 361 diseased cases 137 receive vaccine (37.9%), outcome of case did not need hospital admission, complete recovery for a protective effectiveness of 61% (95% confidence interval [P =.03). overall protective effectiveness of influenza vaccine was 59.3% in children and 66.5% in adult so over all protection 63%, recommendation Routine annual influenza vaccination is recommended for all persons age 6 months and older.

Evaluation of two independent protocols for the extraction of DNA and RNA from different tissues of sea cucumber Isostichopus badionotus

Isostichopus badionotus is a sea cucumber species of great ecological and economic relevance for Mexico and Central American and Caribbean countries; however, the protocols for the extraction of the nucleic acids have not yet been published. In this study, we describe the first protocols to obtain DNA and RNA from different tissues of I. badionotus, which include the respiratory tree, gonad, longitudinal muscle bands, anterior intestine and cloaca. The extraction of high-quality DNA was performed using the DNeasy Blood & Tissue kit (Qiagen, Valencia, CA, USA) with minor modifications in different points of the protocol.Concerning the RNA, the method of TRIzol was used. This method is particularly advantageous in situations where cells or tissues are enriched for endogenous RNases or when the separation of cytoplasmic RNA from nuclear RNA is impractical. The methodologies used in this study allowed us to obtain DNA and RNA of high quality and integrity in the different tissues of I. badionotus, which will be the basis for future genomic and transcriptomic studies.•The successful extraction of DNA and RNA was achieved in the different tissues of I. badionotus.•The concentrations of DNA and RNA obtained were adequate for a diversity of analyses at a molecular level.